Production of Transgenic Mice
The transgenic mice are produced by microinjection of DNA directly into one of the pronuclei of mouse-fertilized zygotes. The transgenic facility provides the service for generating transgenic mice for investigators. We commonly used F2 of C57BL/6 x DBA/2 and FVB/N as for the donor embryo. Other genetic background strains can also be used and injected. Typically, 150~200 embryos will be injected and implanted, which representing two injection days work. The survived pups will be screened by investigators for the presence of the transgene by PCR or southern blot genotyping assays. A minimum of two transgenic founders is guaranteed, or up to a maximum of 200 embryos injected.
The purity of the DNA to be injected is one of the most essential factors affects the successful outcome of pronuclear microinjections. We strongly suggest investigator provide us high purity of DNA for injection. Recommended protocols are available for DNA preparation and purification.
Protocol for Maxi prep and DNA fragement purification
Production of Gene Targeting Animal
Chimeric mice are produced by injecting embryonic stem cells into the cavity of a mouse blastocyst. The most commonly used embryonic stem cells are derived from agouti 129/SV mice and host blastocysts are harvested from black C57Bl/6 mice. The resulting chimeric mice from injected blastocysts will be composed of tissue derived from the inner cell mass of the host blastocyst as well as the embryonic stem cells. It is possible for embryonic stem cells to contribute to all type of cells of the chimeric mouse, including the germ cells.
TG facility offers the microinjection of ES cells (investigator provided) to blastocysts, and implantation services to investigators at the UCLA as well as extramural investigators. Three type services are offered at this moment:
- Basic I service: Implant 50 or more injected blastocysts, which representing one injection day work
- Basic II service: Produce 3 or more chimeric mouse or implant 150 injected blastocysts, representing three injection days of work.
- Premium service: Generate germline transmission line.
The investigator must provide embryonic stem cells proven to be pathogen free. A form to complete before cell submission is found on our order form.
If your ES cells are in the ES cell facility, the staff at ES facility will prepare your cells for injection. Or if you prepare your own cells, we recommend you to follow the linked protocol to prepare cells.
Protocol for Preparation of ES cells for injection
Additional Services:
We also offer embryo collection, cryopreservation, strain rederivation, and in vitro fertilization
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